Hi researchers,
I want to measure IL-6 secreted by cultured cells at different time point, using the supernantant. For example 12, 24, 48h after stimuli. But one thing I realize is that the ELISA kit is 96-well plate and I want to do all the samples at the same time. Then how should I keep the supernatant? Will the IL-6 degrade during the storage?
Or I should do one portion at each time point? This will warm up and cool down the reagents for several times, which I'm not sure whether it will affect the result. Also if I do in separate times, I need generate standard curve every time, which means I will "waste" several wells for the standard curve.
Anyone have experience in this? Thanks!
Hi' you can collect the supernatant and store at -4 for upto 2 month.once collect all sup, you can perform the experiment. Don't to worry, the supernatant not degrade in the - 4 refrigerator. long time store -20 .
Hi I did checks on this. Depending on culture medium composition, IL-6 decreased at 20 C, 4 C and -20 C (probably due to degradation), but my -80 C and liquid nitrogen freezing samples retained their levels for up to 1 year. Suggest you do minimum -80 C freezing - you must aliquot your sample so that you only thaw it once when you assay.
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